High-sensitivity gradient semi-quantitative immunochromatography detection test strip and detection method

Abstract

The invention relates to a high-sensitivity gradient semi-quantitative immunochromatography detection test strip. The test strip comprises test paper and a sample cup; the test paper comprises a supporting layer, an adsorption layer and a protective layer; the adsorption layer sequentially comprises a sample pad, a conjugate pad, a cellulose membrane layer and a water absorption material layer; a detection line T1, a detection line T2 and a quality control line are arranged on the cellulose membrane layer; the conjugate pad is made of glass fiber cotton adsorbed with a nano material labeled antigen; the sample cup is a container for diluting a sample, and a target monoclonal antibody diluted by a gold-labeled protein resuspension is pre-fixed in the sample cup. According to the test paper detection method, the two detection lines which do not interfere with each other can be prepared, and gradient semi-quantification is realized; the antibody is independently dried in the sample cup, so that the antibody is prevented from being folded and damaged in the labeling process, and the dosage of the antibody can be accurately controlled; the nano-particle labeled artificial antigen is used as a probe, each antibody intercepted on the detection line T2 can be accurately combined with two labeled antigens, so that signal self-amplification is realized; and compared with a traditional method, the method is higher in sensitivity.

Description

technical field [0001] The invention relates to an immunochromatography method, in particular to a highly sensitive gradient semi-quantitative immunochromatography detection test strip and a detection method. Background technique [0002] Immunochromatography is a rapid immunoassay technology developed in recent years. Its basic principle is the interaction between antibodies and antigens. It uses nitrocellulose membrane as a carrier to label antigens or antibodies as a tracer. A chromatographic detection technique. Compared with traditional immunoassay methods, immunochromatography has the advantages of strong specificity, accurate detection results, simple equipment operation, rapid measurement, low cost, no need for skilled technicians or expensive equipment, etc., fully meeting the requirements of instant detection . At present, immunochromatography technology has been widely used in medicine, animal husbandry, agriculture and food safety and other fields. [0003] In...

AI Technical Summary

Problems solved by technology

[0003] In the field of food safety, small molecule antigen immunochromatography based on the principle of competition is the most widely used. However, the existing detection mode often encounters that the sensitivity and affinity of antibodies are very good, but the sensitivity of the established immunochromatography test paper is far inferior to that of the established ELISA. The detection method is good, or the color of the detection line of the test paper is weak, etc.

The main reasons are as follows: 1. The antibody folded during the labeling process, and an excessive amount of antibody was added to stabilize the labeling material; 2. The efficiency of the artificial antigen intercepting antibody was low, and an excessive amount of labeled antibody had to be added in order to improve color development. Improve color development; 3. There is no signal amplification when colloidal gold is used to label antibodies. Fluorescent materials such as quantum dots achieve signal amplification while requiring detection instruments to judge the results, which complicates detection.

The existing small molecule immunochromatography detection mode cannot solve this problem, so a feasible and sensitive test strip detection mode is urgently needed to provide more methods for the development of immunochromatography test strips

[0004] At the same time, there is another problem in the detection of small molecules: many targets require different limit standards in different samples, or require gradient semi-quantitative, traditional immunochromatography test paper mode provides only one detection line under naked eye observation to achieve semi-quantitative Quantitatively, some scholars use artificial antigens of different concentrations to spray multiple detection lines to achieve gradient semi-quantitative, but the mutual influence between detection lines is very large, and it is difficult to realize gradient semi-quantitative, so there is an urgent need to prepare gradient semi-quantitative immunoassays that do not interfere with each other. Chromatographic method

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